Objective: To investigate the chemical constituents from the chloroform and ethyl acetate fractions of ethanol extract from Hemistepta lyrata. Method: The compounds were isolated by silica gel chromatography

ODS chromatography

Sephadex LH-20 chromatography

and preparative HPLC

and their structures were elucidated on the basis of chemical and spectroscopic methods

including MS

1D and 2D NMR spectral techniques. Result: Eight flavonoids were isolated from the chloroform and ethyl acetate fractions of ethanol extract from H.lyrata

and were identified as acacetin(1)

cirsimaritin(2)

apigenin(3)

kaempferol(4)

kaempferol-3-O-β-D-glucopyranoside(5)

kaempferol-3-O-β-D-glucopyranoside(6)

kaempferol-3-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside(7)

and acacetin-7-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside(8). Conclusion: This is the first report of the isolation of 5 from Hemistepta genus.